WO1998044972A2 - Dispositif pour isoler des particules, notamment des cellules - Google Patents

Dispositif pour isoler des particules, notamment des cellules Download PDF

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Publication number
WO1998044972A2
WO1998044972A2 PCT/DE1998/001007 DE9801007W WO9844972A2 WO 1998044972 A2 WO1998044972 A2 WO 1998044972A2 DE 9801007 W DE9801007 W DE 9801007W WO 9844972 A2 WO9844972 A2 WO 9844972A2
Authority
WO
WIPO (PCT)
Prior art keywords
capillary
cannula
vessel
cell
manipulator
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/DE1998/001007
Other languages
German (de)
English (en)
Other versions
WO1998044972A3 (fr
Inventor
Klaus-Michael Debatin
Christian Beltinger
Gerd RÜHLE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Deutsches Krebsforschungszentrum DKFZ
Original Assignee
Deutsches Krebsforschungszentrum DKFZ
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Deutsches Krebsforschungszentrum DKFZ filed Critical Deutsches Krebsforschungszentrum DKFZ
Publication of WO1998044972A2 publication Critical patent/WO1998044972A2/fr
Publication of WO1998044972A3 publication Critical patent/WO1998044972A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
    • B01L3/0217—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids of the plunger pump type
    • B01L3/022—Capillary pipettes, i.e. having very small bore

Definitions

  • the invention relates to a device for isolating particles, in particular cells, with a capillary for transporting the particle or particles.
  • the relevant cell is first released using the microdissection and then using the relevant cell
  • Fine steel cannulas are particularly suitable for microdissection.
  • transfer of the released cell with steel cannulas into a vessel is hampered by adhesive or electrostatic forces, and repeated, time-consuming attempts are usually required to attach the cell to the needle.
  • the needle is moved from the object glass in the vicinity of the reaction vessel, the cell is often lost and when the cell is inserted into the reaction vessel, electrostatic forces often push the cell off the needle.
  • Steel cannulas are also unsuitable for aspiration of cells due to the thickness of the wall, the length of the tip and the size of the opening.
  • glass capillaries are very well suited for the aspiration of microdissected cells. Because of their fragility, they are only very suitable for microdissection. Inserting the glass capillary with the aspirated cell into a reaction vessel to eject the cell often leads to the loss or contamination of the cell and is very time-consuming.
  • the invention has for its object to develop a generic device that shortens the time required for cell extraction and is easy to use.
  • the object is achieved in particular with a generic device in which a cannula is arranged around the capillary, in which the capillary can be displaced along its longitudinal axis.
  • Guiding the capillary in a cannula has the advantage that the cannula can be made of a firmer material and is used for microdissection.
  • the fine capillary is guided in a protected manner in the cannula and can also be used for microdissection or to support it, and secondly the capillary is used to hold the particle or cell.
  • the displaceability of the capillary in the cannula allows the cannula to be used exclusively for microdissection when the capillary is withdrawn.
  • the capillary which is moved out towards the front can also be used for microdissection. However, it primarily serves to aspirate the particle.
  • capillary and cannula described thus connects the micro-dissection needle and aspiration capillary and allows both work steps to be carried out with a single device.
  • the capillary can be rotated about its longitudinal axis in the cannula.
  • the often irregular opening of the capillary can thus be placed on a dissected cell in such a way that its aspiration into the capillary is effortless succeed.
  • the production of glass capillaries and metal cannulas has proven particularly useful in practice.
  • the extremely fine glass capillary is protected by the harder metal cannula with its larger diameter.
  • the metal cannula has a front end with a tip.
  • one end of the capillary opens directly into an essentially closed vessel. This allows the cell or any particle to be conveyed into the capillary in the capillary after it has been taken up into the capillary.
  • the closed vessel has a lid element with a capillary opening and a pneumatic opening.
  • the capillary opening serves to introduce the capillary into the vessel and the pneumatic opening to create a negative pressure in the vessel through which the particle can be sucked into the vessel with a drop of liquid.
  • a simple operation of the device is achieved with a manipulator that holds the cannula, the capillary on the same manipulator along the
  • the capillary can be axially displaced relative to the cannula in order to move the capillary out of the cannula to the dissected object.
  • This manipulator is advantageously designed in such a way that the cannula and the capillary are arranged together by means of the manipulator so as to be pivotable about an axis. This allows the capillary and cannula to be easily and quickly guided into the field of view of a microscope and removed from the field of view again.
  • a simple construction of the micromanipulator is achieved in that the axis, about which the cannula and capillary are arranged pivotably, is arranged essentially vertically.
  • the manipulator is made considerably easier by a positioning device which enables precise positioning in the field of view and focus of a microscope above an object. It is particularly advantageous if the positioning device has a latching device which can be adjusted in such a way that it allows simple, quick, rough adjustment of the entire device.
  • FIG. 1 shows a perspective illustration of a receptacle with capillary and cannula
  • FIG. 2 is a perspective view of a micromanipulator with a receptacle, capillary and cannula
  • FIG. 3 schematically shows the handling of the device according to the invention in 3 steps (1st embodiment)
  • FIG 4 shows schematically in 3 steps the handling of the device according to the invention (2nd embodiment).
  • FIG. 1 shows the device 1 without a manipulator 2 (see FIG. 2). It consists of the glass capillary 3, which is guided in a steel cannula 4 and with one end 5 extends into a vessel 6.
  • the vessel 6 is essentially closed and has only one capillary opening 7 through which the capillary ins
  • Vessel is guided, and a pneumatic opening 8 through which an angle tube or a hose 9 can be connected to the vessel.
  • Capillary opening 7 and pneumatic opening 8 are arranged in a cover element 10 which can be connected in a gas-tight manner to a hollow body part 11.
  • an end cover 1 2 is provided, which can be placed on the cover part 10 in such a way that it covers the capillary opening 7 and the pneumatic opening 8.
  • the cannula 4 has a front end 1 4 with a tip 1 5 and at its opposite end 1 6 a perforated plastic plug 17 is fastened, in which the capillary 3 runs.
  • FIG. 1 The device parts shown in Figure 1 are in a micromanipulator 2 - see. Figure 2 - held.
  • This micromanipulator 2 consists essentially of a foot 1 8 to which an arm 19 is rotatably attached.
  • the foot 1 8 is attached to the countertop of an inverted microscope and has a vertical axis 20, about which is fixed to a disk 21 by means of a knurled nut.
  • Stigt arm 1 9 is rotatably mounted.
  • a mandrel 22 On the underside of the disk 21, a mandrel 22 is shown enlarged, which can be snapped into a groove 23.
  • the plastic plug 17 to which the cannula 4 is fastened is inserted into the arm 19.
  • the vessel 6 serving as a cell trap with the capillary 3 is around the
  • a vessel holder 26 is rotated about the axis 24 using a knurled nut 25.
  • a fine drive 27 is arranged between the vessel holder 26 and the arm 19, which allows the container 6 and thus the cannula 3 to be displaced along the axis 24 with a screw 28.
  • the glass capillary 3 which in practice has an outer diameter of 80 ⁇ m and an inner diameter of 40 ⁇ m, is first arranged in the steel cannula 4 in such a way that its front end 29 is completely inside the cannula 4.
  • the cannula 4 has an inner diameter of approximately 1,550 ⁇ m.
  • a small drop of liquid 30 at the front end 29 of the glass capillary 3 makes it easier to later take up a cell 31.
  • the capillary 3 is advanced by means of the screw 28 through the fine drive 27 of the micromanipulator so far inside the cannula 4 until the front end 29 of the glass capillary 3 is in the vicinity of the detached cell or cells. If necessary, the glass capillary 3 is rotated about its longitudinal axis 24 by means of the knurled nut 25, so that its opening is located directly on the cell (FIG. 3b). By suction on Tube 9 of the vessel 6, the cell 31 is aspirated into the glass capillary 3 ( Figure 3c).
  • the glass capillary 3 can also be extended initially in order to microdissect it. This is due to the stable and protective
  • the cannula with capillary is pulled away from the preparation and the manipulator arm 19 is pivoted away to the side.
  • the vessel is connected via the hose 9 to a manual piston pump, by means of which a vacuum can be generated in the vessel 6, which exerts a suction in the direction of the arrow 32 on the liquid drop 30 with the mobilized cell 31.
  • a vacuum can be generated in the vessel 6, which exerts a suction in the direction of the arrow 32 on the liquid drop 30 with the mobilized cell 31.
  • the capillary 3 and the vessel 6 can be centrifuged at 4,000 rpm in a table centrifuge after the cannula has been removed.
  • the cell is then covered in oil at the bottom of the reaction vessel 6.
  • the vessel 6 is removed from the cover element 10, closed with the end cover 1 2 and supplied for further use.
  • the micromanipulator 2 is attached to the work surface of an inverted microscope.
  • the slight pivoting of the arm 1 9 about the axis 20 facilitates loading with the vessel 6, the capillary 3, and the cannula 4.
  • the locking mechanism 22, 23 is set so that the tip 1 5 of the cannula 4 is precisely positioned when it is locked in the field of view slightly above the object and in focus. As a result, the tip does not have to be laboriously re-inserted into the device each time it is swung in after loading or emptying the device
  • the glass capillary opens into a pneumatic tube and a reaction vessel slide is attached to the micromanipulator arm (Fig. 4). With the reaction slide retracted, dissection and aspiration is unhindered (FIG. 4a). The micromanipulation arm is then swung out, the reaction vessel slide extended and a reaction vessel clamped (FIG. 4b). Without touching the capillary and cannula, the reaction vessel slide with the reaction vessel can now be pushed back and the cell after swiveling the
  • Manipulatorarmes are ejected pneumatically into the reaction vessel under microscopic control (Fig. 4c).
  • the device is particularly suitable for the molecular diagnosis of cancer and hereditary diseases on the basis of cytology and histology preparations. It also makes molecular diagnostics from morphological preparations practicable for routine clinical laboratories. The exact determination of the findings plays an important role in particular because the morphological findings lead to far-reaching consequences for the patient, such as surgery, chemotherapy or radiation. Additional, objective parameters are therefore required to assess histological and cytological specimens. Molecular analysis lends itself as such. For such analyzes, cells have to be removed from a histological or cytological preparation. With the device according to the invention it is now possible not to destroy the preparation, which in most cases consists of only a few cells, and nevertheless to be able to make further analysis, for example by means of PCR, accessible.
  • Another area of application of the device according to the invention is
  • fetal cells e.g. erythroblasts
  • the device is useful for obtaining single oocytes, spermatids, blastocysts or embryo cells for pre-implantation diagnosis in in-vitro fertilization in humans and in the context of animal breeding.

Landscapes

  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne un dispositif pour la microdissection et l'aspiration de cellules, dans lequel un capillaire en verre est guidé dans une canule en acier à l'intérieur de laquelle il peut coulisser le long de son axe longitudinal. De préférence, le capillaire en verre et la canule sont guidés avec un seul micromanipulateur. Ce dispositif permet de faciliter considérablement l'obtention de cellules et de réduire le temps nécessaire à cette opération à quelques minutes. Il permet d'utiliser le diagnostic moléculaire à partir de préparations morphologiques même pour des laboratoires cliniques effectuant des examens de routine.
PCT/DE1998/001007 1997-04-10 1998-04-08 Dispositif pour isoler des particules, notamment des cellules Ceased WO1998044972A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19714987.1 1997-04-10
DE19714987A DE19714987C1 (de) 1997-04-10 1997-04-10 Vorrichtung zur Isolation von Partikeln, insbesondere von Zellen

Publications (2)

Publication Number Publication Date
WO1998044972A2 true WO1998044972A2 (fr) 1998-10-15
WO1998044972A3 WO1998044972A3 (fr) 1998-12-30

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1998/001007 Ceased WO1998044972A2 (fr) 1997-04-10 1998-04-08 Dispositif pour isoler des particules, notamment des cellules

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DE (1) DE19714987C1 (fr)
WO (1) WO1998044972A2 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE20304533U1 (de) * 2003-03-21 2004-08-05 Impella Cardiosystems Ag Einführvorrichtung zum Einführen eines Gegenstandes in ein Körpergefäß
DE102007048409A1 (de) * 2007-10-09 2009-04-16 Carl Zeiss Microimaging Gmbh Verfahren zum Positionieren von biologischen Proben in einer mikroskopischen Anordnung
EP2083257A1 (fr) * 2008-01-25 2009-07-29 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt GmbH Procédé et dispositif de transmission d'un échantillon microscopique isolé, système de microdissection doté d'un tel dispositif et procédé de fabrication d'un nanoaspirateur
US8283132B2 (en) 2004-11-05 2012-10-09 Fraunhofer-Gesellschaft Zur Foerderung Der Angewandten Forshung E.V. Method and devices for treating individual biological cells
WO2017061387A1 (fr) * 2015-10-07 2017-04-13 学校法人早稲田大学 Système de prélèvement d'échantillon

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DE19932032C2 (de) * 1999-07-09 2003-07-24 Eppendorf Ag Vorrichtung zur Mikro-Dissektion von Gewebe
DE10307487A1 (de) * 2003-02-21 2004-09-09 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Verfahren und Vorrichtungen zur verletzungsfreien Bewegung einer Sonde durch biologisches Zellmaterial
DE202005000844U1 (de) * 2005-01-18 2005-04-14 Passow, Harald Versuchsanordnung zur Untersuchung der Wirkung von medizintechnischen Laserinstrumenten auf biologische Präparate
DE102007036150A1 (de) * 2007-08-02 2009-02-05 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Verfahren und Vorrichtung zur Aufnahme von biologischen Zellen aus einer Stammzellenkultur
EP2378342A1 (fr) 2010-04-15 2011-10-19 Mmi Ag Système de micromanipulation doté d'un dispositif de protection pour capillaires
EP3274690A1 (fr) 2015-03-24 2018-01-31 Cytotrack APS Procédé et appareil permettant de fixer, détecter et récupérer une cellule individuelle sur une surface

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US3641823A (en) * 1970-04-10 1972-02-15 Rano J Harris Sr Injection device
US3869068A (en) * 1974-06-06 1975-03-04 Hyperion Inc Diluter probe assembly
AU8768175A (en) * 1975-01-09 1977-06-23 Nat Medical Products Corp Hypodermic cannulae
DE8222222U1 (de) * 1982-08-06 1982-11-25 Mehler, Doron, Dr., 3000 Hannover Katheterset
WO1985000314A1 (fr) * 1983-06-30 1985-01-31 Institut Biokhimii I Fiziologii Mikroorganizmov Ak Procede et dispositif permettant d'effectuer des micro-operations sur des cellules
US4772264A (en) * 1986-06-23 1988-09-20 Regents Of The University Of Minnesota Catheter introduction set
DE3718066A1 (de) * 1987-05-29 1988-12-08 Zeiss Carl Fa Verfahren zur mikroinjektion in zellen bzw. zum absaugen aus einzelnen zellen oder ganzer zellen aus zellkulturen
GB8718232D0 (en) * 1987-07-31 1987-09-09 Manchester Inst Science Tech Sampling of material
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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE20304533U1 (de) * 2003-03-21 2004-08-05 Impella Cardiosystems Ag Einführvorrichtung zum Einführen eines Gegenstandes in ein Körpergefäß
US8283132B2 (en) 2004-11-05 2012-10-09 Fraunhofer-Gesellschaft Zur Foerderung Der Angewandten Forshung E.V. Method and devices for treating individual biological cells
DE102007048409A1 (de) * 2007-10-09 2009-04-16 Carl Zeiss Microimaging Gmbh Verfahren zum Positionieren von biologischen Proben in einer mikroskopischen Anordnung
DE102007048409A8 (de) * 2007-10-09 2009-11-05 Carl Zeiss Microimaging Gmbh Verfahren zum Positionieren von biologischen Proben in einer mikroskopischen Anordnung
US8228499B2 (en) 2007-10-09 2012-07-24 Carl Zeiss Microimaging Gmbh Method for positioning biological samples in a microscopic arrangement
EP2083257A1 (fr) * 2008-01-25 2009-07-29 Helmholtz Zentrum München Deutsches Forschungszentrum für Gesundheit und Umwelt GmbH Procédé et dispositif de transmission d'un échantillon microscopique isolé, système de microdissection doté d'un tel dispositif et procédé de fabrication d'un nanoaspirateur
WO2009092495A1 (fr) * 2008-01-25 2009-07-30 Helmholtz Zentrum München Deutsches Forschungszentrum Für Gesundheit Und Umwelt Gmbh Procédé et dispositif pour transférer un échantillon microscopique isolé, système de microdissection doté d’un tel dispositif, et procédé de fabrication d’un nano-aspirateur
US8573073B2 (en) 2008-01-25 2013-11-05 Helmholtz Zentrum Muenchen Deutsches Forschungszentrum Fur Gesundheit Und Umwelt Gmbh Method and device for transferring a microscopic, isolated sample
WO2017061387A1 (fr) * 2015-10-07 2017-04-13 学校法人早稲田大学 Système de prélèvement d'échantillon
JPWO2017061387A1 (ja) * 2015-10-07 2018-08-09 フロンティアバイオシステムズ株式会社 試料採取システム

Also Published As

Publication number Publication date
WO1998044972A3 (fr) 1998-12-30
DE19714987C1 (de) 1998-09-24

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